Automated Cell Counter Research Articles

An Accessible Automatic Cell Counting Methodology Using Trainable Weka Segmentation in ImageJ Validated on Murine Microglia

Abstract Counting cells is a cornerstone of tracking disease progression in neuroscience. A common approach for this process is having trained researchers individually select and count cells within an image, which is not only difficult to standardize but also very time-consuming. We present an automatic cell counting methodology which leverages the Trainable WEKA Segmentation (TWS) plugin available in the FIJI distribution of ImageJ (Schindelin 2012, Schneider 2012). TWS provides access to machine learning tools for object segmentation with an accessible graphical user interface. However, to analyze whole datasets requires further automation through the use of macros which we constructed within ImageJ maintain this high level of accessibility. We compared our automatic counts to a published, hand counted dataset of Immunofluorescence stained mouse cortex microglia (Singh et al. 2020). For this analysis we used a wild-type (WT) control and a transgenic mouse model for HIV-induced brain injury (HIVgp120). HIVgp120 mice showed behavioral deficits, and increase in microglia numbers presumably caused the gp120 protein. 10X images were collected in the sagittal plane from brain slices of three 11–14 month-old male mice for each genotype. The auto counting strategy with TWS replicated the increase in mean microglial density in images from the HIVgp120 mice (Manual p =0.0001; Auto p=0.0002). The auto count also correlates significantly with the manual counts in both genotypes (WT p= 2.297 e−07, adj. R2= 0.65; HIVgp120 p=2.591 e−04 adj. R2=0.423). The auto counting strategy took less than a fifth of the time for the expert manual count, allowing for the efficient exploration of large sets of image data.

Prevalence of anemia and associated factors among the elderly population in South Khorasan, Birjand, 2019.

Background: Anemia is a multifactorial and common public health problem in geriatric age groups, especially in developing countries. Therefore, this study was designed to study the prevalence of anemia and associated factors among the elderly population in Birjand, Iran, in 2019. Methods: This was a cross-sectional approach to the baseline data of the Birjand longitudinal aging study (BLAS) in which 1396 people aged ≥ 60 years were screened for the presence of anemia based on the World Health Organization (WHO) criteria. For each participant, a standard questionnaire was administered. Furthermore, the height, weight, and body mass index (BMI) were calculated. Blood samples were obtained from each participant for hematological examination. Hemoglobin, hematocrit, and other indices of cell blood count were measured using an automatic cell counter. The prevalence rates were estimated using survey analysis with the weight of Birjand county older population. Univariate and multivariable logistic regression analyses were applied to detect the associated factor with anemia. Results: The mean age of the participants was 69.73±7.66 years. The crude prevalence of anemia was 11.10%, and the age-standardized prevalence based on the standard WHO population 2000-2025 was equal to 16.78% (12.81%–21.66%) (15.95% [10.41%–23.69%] in women and 17.32% (12.65%–23.25%) in men. Mild and normocytic anemia were the predominant types. The mean hemoglobin, hematocrit, mean cell volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) were lower in women than in men and the mean platelet count in women was higher (p<0.001). In the final multivariate logistic regression model, only age groups, BMI, fish consumption, and chronic kidney disease (CKD) were related to anemia. Conclusion: In conclusion, our findings showed the association of anemia with some risk factors and diseases. Anemia in geriatric age groups is often underdiagnosed; hence, identification of subgroups at risk for anemia and its associated risk factors in geriatric groups has a paramount importance in preventing adverse outcomes.

Effect of Extra-mammary Diseases on Udder Health and Biochemical Changes in Crossbred Cows with Subclinical Mastitis

Background: Infectious diseases and metabolic disorders are common in crossbred cows and adversely affect optimum production as well as quality of milk. The quality of milk plays a significant role in the production of high-quality dairy products. High somatic cell count (SCC) in the milk significantly decreases the potential of such milk for the production of high quality dairy products. In this study, it is hypothesized that the extra-mammary infections and metabolic disorders increase probability of intra-mammary infections thereby increasing somatic cell count in the milk making it unsuitable for dairy industry.Methods: Cows were grouped based on the diseases or disorders like Teat Stenosis (Gp SCM-TS), Ruminal Acidosis (Gp SCM-RA), Nonspecific Diarrhea (Gp SCM-ND), Respiratory Tract Infections (Gp SCM-RTI) and Repeat Breeder Syndrome (Gp SCM-RD). Diagnosis of subclinical mastitis was made on the basis of California Mastitis Test (CMT) scores and by Somatic Cell Count (SCC) by automatic somatic cell counter. Biochemical parameters analyzed in automatic biochemical analyzer using commercially available kits.Result: The mean SCC values significantly higher in cows with sublinical mastitis and with concurrent infectious and metabolic diseases. Similarly, the concentrations of serum alkaline phosphatase (ALP), aspartate amino transferase (AST), and alanine amino transferase (ALT) were higher in affected cows. Concentrations of serum total proteins (TP) and blood urea nitrogen (BUN) in all the groups of affected animals were higher. The changes in the calcium (Ca) and phosphorus (P) levels were not observed in cows with subclinical mastitis and with other diseases under study. The SCC values did not correlate with the values of ALP, AST, ALT, TP, BUN, Ca and P among the studied groups. It can be concluded that animals with concurrent infections and metabolic disorders increase the SCC and influence the alteration in the biochemical parameters of subclinical mastitic animals.

Evaluation of morphology and viability of spheroid derived from Insulin-GLase cell line: A model system to understand Type 2 Diabetes Mellitus

Type 2 Diabetes Mellitus (T2DM) is one of the major health issues in the world. The cellular mechanism of T2DM is still not fully understood. It could be studied by using spheroid three-dimensional (3D) culture which is considered representative of the in vivo conditions. Several types of pancreatic β cell lines have been used, one of which is the insulin-GLase (iGL) cell line. This study aims to evaluate the effect of cell density and incubation time on spheroid morphology and cell viability in order to understand which one can be considered as the best option in studying T2DM using iGL cell. Spheroid was made by using the Hanging drop method. The variations of initial seeding cells were 50, 100, 200, and 400 cells/µL then incubated for 1, 2, 3, and 4 days. The evaluated parameters in this study are spheroid morphology and cell viability. Spheroid morphology was observed by using inverted phase contrast microscope integrated with camera (Nikon) and NIS-Elements Analysis D software. Cell viability was determined by using LUNA-II™ Automated Cell Counter (Logos Biosystem). The result of this study showed that spheroid in all of the group cell concentration have formed since the first day and its diameter was significantly increased on the following days (p&lt;0,05). The spheroid size was positively correlated with the cell density in group 50-200 cells/µL. A single and stable spheroid morphology was observed in 50-100 cells/µL group. Cell viability in 3D culture system was lower and significantly decreased since day 3 compared to 2D culture (p &lt;0.05; 0.01). In conclusion, spheroid derived from iGL cell line with a stable morphology and good viability could be obtained from a cell concentration of 50-100 cells / µL with two days of incubation.

“MEAN PLATELET VOLUME CORRELATED TO GLYCEMIC CONTROL IN TYPE 2 DIABETES MELLITUS”

Background: Diabetes mellitus (DM) is becoming a global pandemic. The number of people with diabetes in India increased from 26·0 million in 1990 to 65·0 million in 2016. Diabetes mellitus (DM) has been considered as a 'prothrombotic state' with enhanced platelet reactivity. Diabetic patients have an increased risk of developing micro and macro vascular diseases, and platelets may be involved as putative agents owing to their altered morphology, function and activation. It is an established fact that the value of glycated hemoglobin (HbA1c), a marker of long-term glucoregulation, should be kept below 7% in order to reduce the risk of micro and macrovascular complications in Type 2 Diabetes Mellitus (DMT2) patients. Mean platelet volume (MPV) has been correlated with vascular complications of DMT2. MPV as a marker of platelet size, function &amp; activation may serve as another potential marker of risk of micro-vascular and macrovascular complications in DMT2 patients. Aims-To study correlation between MPV, and HbA1c in DMT2 patients. Methods- Over a period of 24 months, patients aged between 30 to 60 years, diagnosed with DMT2 and subtyped based on American Diabetic Association Criteria (ADA) as having HbA1c either less than 7% - DMT2 (Controlled group) or with HbA1c more than 7% - DMT2 (Uncontrolled group) were included in the present study. Non-diabetic patients ('Non Diabetic Group) were included based on their fasting and post prandial blood glucose levels and served as controls. Venous blood samples were tested for fasting, postprandial and random blood sugar estimation by GOD-POD method, MPV by automated cell counter and HbA1c by HPLC, within one hour of sampling. Results were statistically tested using (R) unpaired t Test by SPSS Software (version 22). Results- Among 236 patients studied, 66 were 'Non-diabetic controls' &amp; 170 were having diabetes (DMT2). Of those having DMT2, 94 patients belonged to DMT2 (Uncontrolled group) 76 patients belonged to DMT2 (Controlled group). MPV in Non-diabetic group, DMT2 (Controlled group) and DMT2 (Uncontrolled group) was 10.01 ± 1.12 , 10.76 ± 1.11  and 11.67 ± 1.83  respectively. MPVwas signicantly higher in DMT2 group compared to Non-diabetic group (10.82 ± 1.31  vs 10.01 ± 1.12 , p &lt; 0.0001), MPV was also signicantly higher in DMT2 (Uncontrolled group) compared to DMT2 (Controlled group) (11.67 ± 1.83  vs. 10.76 ± 1.11 , p &lt; 0.0001). Conclusion- In the present study Mean Platelet Volume was found to be signicantly higher in Diabetes Mellitus Type 2 patient compared to Nondiabetic patients. Higher MPV also correlated with higher HbA1c, being higher in those with higher HbA1c. It can be concluded that MPV may be useful as inexpensive surrogate marker of HbA1c in the diagnosis and prognosis of vascular complications of DMT2 patients.

Automated cell count in body fluids: a review.

Body fluid cell counting provides valuable information for the diagnosis and treatment of a variety of conditions. Chamber cell count and cellularity analysis by optical microscopy are considered the gold-standard method for cell counting. However, this method has a long turnaround time and limited reproducibility, and requires highly-trained personnel. In the recent decades, specific modes have been developed for the analysis of body fluids. These modes, which perform automated cell counting, are incorporated into hemocytometers and urine analyzers. These innovations have been rapidly incorporated into routine laboratory practice. At present, a variety of analyzers are available that enable automated cell counting for body fluids. Nevertheless, these analyzers have some limitations and can only be operated by highly-qualified laboratory professionals. In this review, we provide an overview of the most relevant automated cell counters currently available for body fluids, the interpretation of the parameters measured by these analyzers, their main analytical features, and the role of optical microscopy as automated cell counters gain ground.

Compact and automated particle counting platform using smartphone-microscopy

The majority of existing advanced automated cell counter instruments used in laboratory settings are complex, expensive, and bulky. As a result, applications of these instruments have been limited to such laboratories. Meanwhile, in many rural areas and developing countries, clinical laboratories equipped with optical microscopy, hematology analyzers or commercial automated particle counters may not be readily accessible to everyone. However, in the same regions, the number of cell-phone users are rapidly increasing, suggesting a need to develop an easy-to-use and portable smart-phone based cell counting technology that can be leveraged in resource-limited areas. To this end, we present an automated, portable and easy-to-use smartphone-based particle counting platform designed to detect particles in a sample solution. This novel pump-less, flow-based portable technology utilizes a small lens attached to a smartphone camera to magnify particles passing through a microfluidic channel, and record a video using a smartphone camera application. The captured video is transmitted to a local computer to be processed through a custom-developed algorithm to perform particle counting. Using a total of 30 different test samples, we have shown that our technology can detect and count polystyrene beads as small as 16.2 μm with ~100% accuracy. We evaluated the performance of our new technology using different size particles and showed that the results are comparable to the tedious, manual-microscope based counting method. Although, we benchmark the performance of the platform using polystyrene beads, we emphasize the wide applicability of the platform to a wide range of biological, biomedical, and industrial applications.

Unusual Morphological and Automated Hematology Analyzer Features in 3 Cases of B-cell Malignancy-associated Type I Cryoglobulinemic Vasculitis.

Type I cryoglobulins are monoclonal immunoglobulins produced due to underlying hematological malignancy. Cryoglobulins spontaneously precipitate from serum and plasma at low temperatures and become soluble again on rewarming to 37°C. Processing of blood at temperature lower than 37°C in the laboratory may cause precipitation of cryoglobulins resulting in interferences in the automated cell counter analysis. We report three patients with cryoglobulinemic vasculitis wherein each case had different morphology of cryoglobulin precipitates on peripheral blood film, like needle shaped bluish-gray crystals, amorphous weakly basophilic extracellular deposits extraneously indenting red blood cells and basophilic neutrophilic inclusions respectively. The effect of cryoglobulins on two technologically different automated cell counters based on principles of impedance, Volume-Conductivity-Scatter (VCS) and fluorescence flow cytometry was assessed. This case series provides interesting insight into the varying morphological features of cryoglobulins on May-Grunwald-Giemsa stained blood films and interference caused by cryoglobulins in different automated cell counter analysis resulting in pseudo-leucocytosis, pseudo-thrombocytosis, abnormal histograms and scatterplots. Identification of these hematologic abnormalities and artifacts induced by cryoglobulins is necessary since it may be the first clue leading to the timely diagnosis of cryoglobulinemia and hence the underlying hematological malignancy, as in our cases.

STUDY OF PLATELET VOLUME INDICES IN PATIENTS OF CORONARY ARTERY DISEASE

Background: Various relations have been found out regarding the role of platelets in thrombosis and coronary heart disease. This study is an earnest attempt to delineate the relations between the various platelet volume indices (PVI) and their role in coronary heart disease. Method: During the study period of April 2014 to March 2015, total 180 subjects were studied. Out of them 120 were cases and 60 were controls. The cases of coronary artery disease were divided in to two groups A and B, according to their treatment. Group A included 60 patients with acute coronary syndromes, mainly AMI and UA with medical treatment. Group B included 60 patients with any percutaneous coronary interventions (PCI) for previous ischemic event. Group C included 60 healthy individuals from health check-up unit. A brief and relevant clinical history and laboratory investigations will be taken on the prepared Performa for the subjects. The EDTA and plain samples of 180 subjects were processed in the Central Diagnostic Laboratory. Results: The present study showed higher PVI in the group A and group B when compared to the control group. MPV, PDW and P-LCR were signicantly reduced after treatment in both groups A and B. The mortality was more in those with higher PVI. Conclusion: Coronary artery disease is associated with signicant morbidity and mortality. Platelet volume indices in histograms are easily generated by automated cell counter. Thus, by assessment of it we can predict an impending coronary event. We can prevent reinfarction by monitoring of it. The risk of acute coronary event is also decreased after treatment.

Point-of-Care Testing Effectiveness on Blood Donor Hemoglobin Testing.

Hemoglobin (Hb) evaluation by point-of-care testing (POCT) identifies borderline or anaemic asymptomatic blood donors. Although quality control checks confirm that this device is fit for use, it is still not clear whether the analyser is performing effectively. A protocol comparing the POCT EKF Diagnostics with the Sysmex XN-550 automated cell counter (ACC) has been designed. Various scenarios of Hb measurements from the ACC and the POCT device are compared using the Spearman correlation and Intraclass correlation. The Bland-Altman method was used to analyse the level of agreement between the two devices. Correlation between the two devices was best observed in the venous vs venous blood scenario. The POCT device overestimates the Hb levels in capillary blood, meaning that Hb requirements should be adjusted and when feasible testing repeated on venous blood using an ACC. Furthermore, it is suggested thar each Facility determine their own Hb threshold.

Automated cell differential count in sputum is feasible and comparable to manual cell count in identifying eosinophilia

Introduction Cell differential count (CDC) of induced sputum is considered the gold standard for inflammatory phenotyping of asthma but is not implemented in routine care due to its heavy time- and staff demands. Digital Cell Morphology is a technique where digital images of cells are captured and presented preclassified as white blood cells (neutrophils, eosinophils, lymphocytes, macrophages, and unidentified) and nonwhite blood cells for review. With this study, we wanted to assess the accuracy of an automated CDC in identifying the key inflammatory cells in induced sputum. Methods Sputum from 50 patients with asthma was collected and processed using the standard processing protocol with one drop 20% albumin added to hinder cell smudging. Each slide was counted automatically using the CellaVision DM96 and manually by an experienced lab technician. Sputum was classified as eosinophilic or neutrophilic using 3% and 61% cutoffs, respectively. Results We found a good agreement using intraclass correlation for all target cells, despite significant differences in the cell count rate. The automated CDC had a sensitivity of 65%, a specificity of 93%, and a kappa-coefficient of 0.61 for identification of sputum eosinophilia. In contrast, the automated CDC had a sensitivity of 29%, a specificity of 100%, and a kappa-coefficient of 0.23 for identification of sputum neutrophilia. Conclusion Automated- and manual cell counts of sputum agree with regards to the key inflammatory cells. The automated cell count had a modest sensitivity but a high specificity for the identification of both neutrophil and eosinophil asthma.

The Combination of Jiedu Xiaoluo Decoction with Autologous Peripheral Blood Stem Cell Transplantation (APBSCT) Accelerates Disease Remission of Non-Hodgkin Lymphoma

Objective This study aimed to explore the therapeutic effects of autologous peripheral blood stem cell transplantation (APBSCT) with Jiedu Xiaoluo decoction (JDX) on non-Hodgkin lymphoma (NHL). Method B lymphoma cells A20 were used to establish nude mice-transplanted tumor model. The peripheral blood of mice was analyzed by automatic blood cell counter. Inflammatory cytokines in tumor tissues were measured by ELISA, real-time qRT-PCR, and western blotting assays. Immunohistochemical staining was employed to evaluate tumor cell growth and apoptosis. CCK8 and Transwell assays were used to detect cell viability, migration, and invasion. Cell apoptosis in vitro was evaluated with flow cytometry. Result In the in vitro co-culture system of A20 cells and hemopoietic stem cells (HSC), JDX notably inhibited the proliferation, migration, and invasion and promoted apoptosis of A20 cells compared to HSC treatment alone. In animal tumor xenografts of NHL, the combination of APBSCT with JDX significantly promoted hematopoietic reconstitution, inhibited tumorigenesis of A20 cell, promoted the inflammatory microenvironment remission, inhibited cell proliferation, and promoted apoptosis compared to APBSCT alone. Conclusion The combination of APBSCT with JDX might be an effective strategy to treat NHL through inhibiting tumorigenesis and reconstructing hematopoietic and immune microenvironment. Our finding provided a novel insight into the clinical application of Traditional Chinese Medicine (TCM) against NHL.

The antimalaria drug artemisinin displays strong cytotoxic effect on leukaemia lymphocytes in combination with vitamin C and pro-vitamin K3

This study investigated the anticancer effect of the anti-parasitic drug artemisinin in combination with two redox modulators: vitamin C and pro-vitamin K3 (C/K3) The experiments were conducted on leukaemia cells Jurkat. Cells were treated with either artemisinin or C/K3 alone and with all three compounds. Cell proliferation and viability were analysed using trypan blue stating and automated cell counting. The results showed that artemisinin (&gt;10 mM) suppressed cell proliferation activity, but did not induce cell death up to 500 mM. The drug demonstrated a clear cytostatic effect at concentrations 250- 500 mM – Jurkat cells did not proliferate, but were alive. The combination C/K3 (200:2, 300:3 mM/mM) applied alone did not affect cell proliferation and viability. Vitamins C/K3 in concentration ratio 500:5 (μM/mM) decreased cell proliferation activity by ~10%. The triple combination artemisinin/C/K3 manifested synergistic anti-proliferative effects at all concentration ratios analysed. This synergistic effect increased with increasing C/K3 concentration. Based on literature data, it was assumed that the anti-proliferative effect of the triple combination was mediated by changes in the redox-homeostasis of cancer cells. The C/K3 redox system likely acted on cancer mitochondria and increased superoxide production and activation of pro-apoptotic signals, specific for cancer cells. On the other hand, artemisinin could generate hydroxyl radicals as a result of activation of Fenton reactions, depleting intracellular reducing equivalents. Both redox mechanisms lead to activation of signal pathways for induction of cancer cell death.

Intratumoural immune signature to identify patients with primary colorectal cancer who do not require follow-up after resection: an observational study.

Following surgical and adjuvant treatment of primary colorectal cancer, many patients are routinely followed up with axial imaging (most commonly computerised tomography imaging) and blood carcinoembryonic antigen (a tumour marker) testing. Because fewer than one-fifth of patients will relapse, a large number of patients are followed up unnecessarily. To determine whether or not the intratumoural immune signature could identify a cohort of patients with a relapse rate so low that follow-up is unnecessary. An observational study based on a secondary tissue collection of the tumours from participants in the FACS (Follow-up After Colorectal Cancer Surgery) trial. Formalin-fixed paraffin-embedded tumour tissue was obtained from 550 out of 1202 participants in the FACS trial. Tissue microarrays were constructed and stained for cluster of differentiation (CD)3+ and CD45RO+ T lymphocytes as well as standard haematoxylin and eosin staining, with a view to manual and, subsequently, automated cell counting. The tissue microarrays were satisfactorily stained for the two immune markers. Manual cell counting proved possible on the arrays, but manually counting the number of cores for the entire study was found to not be feasible; therefore, an attempt was made to use automatic cell counting. Although it is clear that this approach is workable, there were both hardware and software problems; therefore, reliable data could not be obtained within the time frame of the study. The main limitations were the inability to use machine counting because of problems with both hardware and software, and the loss of critical scientific staff. Findings from this research indicate that this approach will be able to count intratumoural immune cells in the long term, but whether or not the original aim of the project proved possible is not known. The project was not successful in its aim because of the failure to achieve a reliable counting system. Further work is needed to perfect immune cell machine counting and then complete the objectives of this study that are still relevant. Current Controlled Trials ISRCTN41458548. This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 25, No. 2. See the NIHR Journals Library website for further project information.

Colour thresholding-based automatic Ki67 counting procedure for immunohistochemical staining in meningioma

Nuclei segmentation is the initial process in the histopathological image analysis. This process plays a vital role in cell counting. The Ki67 is a nuclear that was widely used among the pathologists to measure the tumour cell proliferation. Generally, the pathologists use the manual counting technique for counting the Ki67 cells. However, the counting results have poor reliability and lack of accuracy. The current study aimed to propose an automatic Ki67 cell counting for meningioma images by using the colour thresholding approach. The proposed method has been tested on 12 photomicrographs of meningiomas. The results showed that the proposed method was able to segment the positive and negative Ki67 cells with an average accuracy of more than 90%. For counting results, the proposed system produced good results in counting the Ki67 cells with an average relative accuracy of 0.91 for positive Ki67 cells and 0.89 for negative cells.

Comparative Evaluation of Platelet Count in Whole Blood and Injectable Platelet-rich Fibrin

Abstract Background: Platelets are formed from megakaryocytes and platelets are present in the circulation for 5–7 days. Platelets are essential in hemostasis, vascular integrity, angiogenesis, inflammation, innate immunity, and wound healing. To capitalize on the advantageous qualities of platelets, platelet-rich fibrin (PRF) was developed by centrifuging peripheral blood. A new platelet concentrate has recently been developed, by utilizing lower centrifugation speed. This new platelet concentrate is in an injectable form called injectable PRF (I-PRF). Aim: This study intends to quantify and compare the platelets in whole blood (WB) and I-PRF. Materials and Methods: The study included ten systemically healthy individuals. Seven milliliter of blood was collected by following standard aseptic protocol. Smears of blood and I-PRF were made and stained by Leishman's stain to confirm the presence of platelets. I-PRF was made by centrifuging 5 ml of blood at 1000 rpm for 3 min at 60 g, which was fed to the automated cell counter unit to count the number of platelets. Results: The platelet count in I-PRF was significantly more when compared with that of WB. Conclusion: The results of our study suggest that the I-PRF has a richer concentration of platelets when compared to the WB.

Paramountcy of platelet parameters in thrombocytopenia- Our hospital experience

Introduction: Thrombocytopenia defined as platelet count below 1,50,000ul. This fall in platelet count may be due to decreased production, increased destruction and pooling of platelets. The platelet parameters include platelet count, mean platelet volume (MPV), platelet distribution width (PDW) obtained by automated cell counters. These help in knowing the pathomechanism of myriad cases of TCPs. Hence the present study was undertaken to know the variations and correlations of platelet parameters in various causes of TCPs. Materials and Methods: 700 blood samples of patients with platelet count 1,50,000 ul were studied. Hematological analysis was done by Sysmex KX-21. Platelet parameters of all the cases and control group were noted. The cases were grouped into various categories of A, B or C depending on mechanism of decrease in platelets. Data was analyzed and tested for statistical significance using one-way ANOVA and post hoc testing using Tukey’s test. Results: Majority of the cases in the present study were in group A (75.3%), followed by group B (23.9%) and group C (0.6%). A higher value of MPV (10.15±1.30 fl) and PDW (14.44±2.35%) was found in group A compared to other categories. Significant results of MPV and PDW values were seen in group A and group B category. Conclusion: Platelet parameters like MPV and PDW obtained by automated cell counters together with platelet counts help us to know the pathomechanism of various causes of TCPs which inturn provides pivotal information for better management. Keywords: Thrombocytopenia, Platelet count, Mean platelet count, Platelet distribution width.

Anemia in Type 2 Diabetes Mellitus (T2dm) and its association with vitamin B12 deficiency

Introduction: India is the diabetic capital of the world. Vitamin B12 deficiency has been recognized since many years ago as an important side effect in diabetic patients who take metformin for more than 5-10 years.This research aimed to determine the prevalence of anemia in type 2 diabetes mellitus (T2DM) and its association with vitamin B12 deficiency due to use of Metformin. Materials and Methods: Total 200 type 2 diabetes mellitus (T2DM) males and females were included in the study.Blood collection for Fasting blood glucose and Hba1c levels were measured from all participants in tubes without anticoagulant. Hemoglobin, mean corpuscular volume (MCV) were measured by automated cell counters. Measurement of vitamin B12 level was done only in those patients who presented with Macrocytic anemia. Results: Among 200 patients, 113 patients were on vegetarian. The prevalence of anemia in this study population was 65(32.5%). 11(16.9%) were diagnosed as Normocytic Normochromic anemia, 22 (33.8%) with Microcytic Hypochromic Anemia, 26(40%) with Macrocytic Anemia and 6(9.2%) with Dimorphic anemia. Among these 32 patients, who presented with macrocytic and dimorphic anemia, vitamin B12 levels were measured. For 23 vitamin B 12 level was200 pg/ml. 149 patients were on Metformin. Conclusion: Our findings suggest that correction of anemia may have a significant role in prevention of other diabetic complications, thus we recommend that treatment criteria for diabetes should include routine hematological tests and annual screening of Vitamin B12 deficiency along with supplementation should be adopted while treating T2DM. Keywords: Macrocytic Anemia, Type-2 Diabetes Mellitus.

An Intelligent Model for the Detection of White Blood Cells using Artificial Intelligence

Background and Objective: The automatic detection and counting of white blood cells (WBCs) play a vital role in the diagnosis of hematological diseases. Computer-aided methods are prevalent in the detection of WBCs because the manual process involves several complexities. In this article, a complete automatic detection algorithm to recognize the WBCs embedded in cluttered and complicated smear images of blood is designed.Methods: The proposed algorithm uses the ellipse detection approach to approximate the presence of WBCs in the Blood. A newly designed artificial electric field algorithm with novel velocity and position bound (AEFA-C) is employed for this purpose. The problem of detection of WBCs is transformed into an optimization problem where the random candidate solutions (ellipses) are efficiently mapped. These candidate ellipses are mapped onto the edge map of the smear image, and a complete mapping is obtained using the AEFA-C algorithm.Results: The effectiveness of the AEFA-C based detector is tested over the 60 smear images of the blood, having all the five types of WBCs or leukocytes. The developed algorithm obtained an overall detection accuracy of 96.90%. Further, the robustness test is performed on the same dataset which justifies that the technique can handle the different noises with the detection accuracy of 90.33%. Also, the comparative study of the proposed detection algorithm with the state-of-art detection algorithms is carried out.Conclusions: The experimental results demonstrate the efficiency of the proposed scheme for the detection of the WBCs in terms of detection accuracy, stability, and robustness and its outperformance over the state-of-art algorithms.

Blood Cellular Changes Associated with Bacteremia and Malaria Co-morbidity among Children in Western, Kenya

Background: Malaria and bacteremia co-morbidity in children cause changes in blood cellular components. Complete blood count from children whose haemoglobin genotypes and bacteremia tests are not known, greatly influence clinical management and interpretation of the haematology results in resource limited healthcare facilities.&#x0D; Objectives: We investigated cellular components from children with bacteremia and malaria co-morbidity. We also analysed the haemoglobin genotypes and bacteria isolates from children with haemoglobin AA, SS and AS in western Kenya.&#x0D; Methods: A total number of 384 children were recruited and complete blood counts done with an automated cell counter. Microscopy was used to determine malaria infections, while bacteremia was determined by blood culture. The haemoglobin genotypes were analysed using the electrophoresis technique.&#x0D; Results: Children with haemoglobin AA and AS had elevated granulocyte counts. Most of the bacteria isolates were from children with malaria and haemoglobin AS. The bacteria isolated from blood culture included non-typhi salmonella, Escherichia coli, Enterobacter cloacae, Staphylococcus aureus, Listeria monocytogenes, Streptococcus pyogenes and Viridans.&#x0D; Salmonella species and staphylococcus aureus were the most prevalent bacteria isolates associated with bacteremia in children with haemoglobin AS and malaria positive.&#x0D; Conclusion: Children with Hb AS have higher chances of malaria and bacterial co-infection which leads to lymphocytopenia, erythrocytopenia and thrombocytopenia. Bacteria responsible for most of malaria co-infections in this region are Salmonella species and Staphylococcus aureus. The malaria and bacterial co-infection in pre-school children initiate differential cellular changes which should be investigated further.